Rice leaf spot have been observed on many fields during survey of rice diseases in three regions in Madagascar. Curvularia sp. was the most frequently isolated pathogen from the most severe diseased ... [more ▼]

Rice leaf spot have been observed on many fields during survey of rice diseases in three regions in Madagascar. Curvularia sp. was the most frequently isolated pathogen from the most severe diseased samples. This pathogen has been reported worldwide to cause important losses. Then in this work, we study the possibility to develop an alternative method to control Curvularia sp. Plants or plant extracts have been traditionally used to control human and plant diseases. Nowadays, the problem of residues of phytopharmaceutical products, reinforce the need for research on the development of natural plant extracts to control plant diseases. Madagascar has many endemic plants of interest. The antimicrobial properties of essential oils from Malagasy aromatic plants were evaluated. The main steps of this study are (i) the in-vitro screening of 39 Malagasy essential oils to control this pathogen, (ii) the evaluation of the phytotoxicity (on rice plants) of the efficient essential oils and (iii) the in vivo test of the activity of these non phytotoxic essential oils on Curvularia sp. on rice seedlings. The activity of essentials oils on Curvularia sp. has been tested in vitro during a microscale ELISA plate bioassay using optical density for a rapid evaluation of the biofungicidal activity of the extracts. Essential oils that showed interesting antifungal effect on this pathogen were tested for their phytotoxicity on rice plants. Briefly, one to two drop(s) of essential oils were applied on 3 leaves leaves of three weeks rice seedling. Six essential oils over the 39 tested showed an interesting antifungal activity with a percentage of growth inhibition greater than 70%. Three essential oils tested were phytotoxic as they induced necrotic spots on rice leaves after 48 h. Among three essential oils left, one of them showed an important effect when applied on rice seedling inoculated with a conidial suspension of Curvularia sp. These results confirmed that some essential oils present an antifungal activity. And even some of them are phytotoxic on rice plants, some essential oils show interesting potential to be an alternative method against rice diseases. [less ▲]

The Peach latent mosaic viroid (PLMVd) infects peach trees in all production areas. This pathogen is responsible of direct and indirect crop losses. However only a few data are available as regards on one ... [more ▼]

The Peach latent mosaic viroid (PLMVd) infects peach trees in all production areas. This pathogen is responsible of direct and indirect crop losses. However only a few data are available as regards on one hand the determinant of pathogenicity of this viroid and on the other hand the resistance mechanisms of plants against this pathogen. The original approach of this work was to give the foundation of this double characterization. Firstly, the role of the P8 pseudoknot, present in every sequenced PLMVd, was studied by directed mutagenesis. Secondly, the molecular response of different peach trees infected by different variants was evaluated by the use of the cDNA-AFLP. The main objective of this thesis was to identify a metabolic pathway implicated in the plant defence against the PLMVd. In the term of this work, it seemed that the P8 pseudoknot was implicated either in the stability or in the replication of the viroid into the infected cells. Indeed, the inoculated variant (with a destabilized pseudoknot) has shown a reduced replication during the cultural season. In spite of the presence of the mutated variant in the plants, no symptom was observed on the peach tree leaves. However, we cannot conclude if this absence of symptom is due to the low viroid quantity either to an implication of the pseudo-knot in the pathogenicity of the PLMVd. The characterization of the gene expression in the infected peach trees has allowed to highlight that the PLMVd represses genes implicated in the photosynthesis and more specifically genes involved in the protection of the two photosystems. This particular gene expression in the infected leaves was linked to the chlorosis and mosaic induced by the PLMVd. However, we cannot conclude with certitude if these symptoms are a cause or a consequence of this particular genes expression. The cDNA-AFLP has also allowed to identify the repression of genes coding for heat shock proteins (HSPs) in symptomatic leaves. These proteins generally have a role in the protein folding, assembly, translocation, stabilization and degradation. The regulation of their expression may have a great influence in the infected plants and, maybe, play a role in the symptoms expression. The gene coding for the novel cap-binding protein (nCBP) was also identified has repressed in the symptomatic leaves. The biological role of these proteins is unclear but it seems that these proteins act in the regulation of the mRNA translation. The repression of nCBP may thus have an important impact and to destabilize various biological pathways. Finally, two genes implicated in the plant defence were identified. One coding for a polygalacturonase inhibitor (over-expressed in symptomatic leaves) and the other one coding for a thiamine intermediate (involved in the SAR and over-expressed in the non-symptomatic leaves). The role of these proteins in the plant defence against the PLMVd is however unclear. To our knowledge, this is the first work where the host-pathogen relationship established during a PLMVd infection are studied. This is also the first time were the gene expression is linked to the viroid-induced symptoms. [less ▲]

Viroids are plant pathogens infecting a broad range of herbaceous and tree crops. Among them, the Peach latent mosaic viroid (PLMVd) infects mainly peach trees, causing a loss of production with no ... [more ▼]

Viroids are plant pathogens infecting a broad range of herbaceous and tree crops. Among them, the Peach latent mosaic viroid (PLMVd) infects mainly peach trees, causing a loss of production with no curative options. Detecting this viroid is thus important for certification procedures aiming to avoid the release of infected material into orchards. Presented here is a complete detection method based on reverse transcription (RT) followed by a quantitative real-time polymerase chain reaction (PCR). New primers were selected and optimal reaction conditions determined for routine application of the method. The technique is 105 times more sensitive than the endpoint RT-PCR used for PLMVd detection, and permits earlier detection of PLMVd in infected plants. The quick, low-cost extraction procedure used and the quality of the results obtained make this method suitable for routine testing. [less ▲]

Viroids are non encapsidated small RNA plant pathogens unable to produce any protein. They are able to infect dramatically a broad range of plants including herbaceous and tree crops. The ways by which ... [more ▼]

Viroids are non encapsidated small RNA plant pathogens unable to produce any protein. They are able to infect dramatically a broad range of plants including herbaceous and tree crops. The ways by which viroids are able to induce diseases are actually unknown. However, recent studies have shown that viroids are able to regulate the gene expression of their hosts, they can modify the host-protein phosphorylation sensibility and they interact with host-protein implicated RNA trafficking and protein phosphorylation. Moreover during their evolution plants have developed a mechanism able to regulate their gene expression and to degrade exogenous RNAs like viroids: the gene silencing. Unfortunately, this pathway seems, now, also highly implicated in the symptoms development. This review describes studies that are realized since a few years to increase the knowledge about the plant-viroid relationship. [less ▲]

Since several years, the Plant Pathology Unit studies the Peach latent mosaic viroid (PLMVd) and other viroids from fruit trees. These studies have led, first, to the lightening of new pseudoknot ... [more ▼]

Since several years, the Plant Pathology Unit studies the Peach latent mosaic viroid (PLMVd) and other viroids from fruit trees. These studies have led, first, to the lightening of new pseudoknot interactions potentially involved in the pathogenic mecanisms of the PLMVd. Secondly the existence of recombination events during the replication of the viroids was expressed. The first goal of this work was to create a collection of dimeric variants to be able to verify these hypothesys during the following years. Dimeric constructions of severe and latent variants were successfully obtained. Unfortunately, the variants selected to perform the recombination analysis contained too much mutations to be used. Finally two design of specific primers were performed. The first provides the possibility to create mutations in the pseudoknots and the second makes us able to detect recombinant variants. The second goal of this work was to set up the cDNA-AFLP on peach extracts to perform the caracterization of host-pathogen interactions between peach trees and the PLMVd.This will be made with collaboration of the Biochemistry and Catalityc RNA Departement of the Universtiy of Sherbrooke wich will analyze the miRNA pattern of infected and healthy plants. Based on former pathogenicity studies of differents variants performed in our Departement, the comparison of differentially expressed genes will include symptomatic and non symptomatic plants and also an healthy plant. Total RNA extracts were successfully performed and a first AFLP profile was obtained. However some steps of the manipulation need an optimization to produce reliable results. [less ▲]